LOCUS       sequence-273584-       10966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein purification of Himar-dCas9 with 3x-FLAG, SV40 NLS, and 
            6x-His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10966)
  AUTHORS   Chen SP, Wang HH
  TITLE     An Engineered Cas-Transposon System for Programmable and 
            Site-Directed DNA Transpositions.
  JOURNAL   CRISPR J. 2019 Dec;2(6):376-394. doi: 10.1089/crispr.2019.0030. Epub
            2019 Nov 19.
  PUBMED    31742433
REFERENCE   2  (bases 1 to 10966)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1089/crispr.2019.0030"; journalName: "CRISPR J"; date: 
            "2019-12"; volume: "2"; issue: "6"; pages: "376-394"
COMMENT     Sequence Label: pET-Himar-dCas9-mammalian
FEATURES             Location/Qualifiers
     source          1..10966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     209..228
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        209..227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    228..252
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             267..289
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             299..364
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             371..391
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1493..1519,1520..1522,1523..4009,4010..4012,
                     4013..5596)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5597..5614
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5667..5685)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5681..5728
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5966..5984)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6052..6156
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6157..6225,6226..7017)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6375..6394)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7188..7776
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7677..7696
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7930..7947
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7962..8101
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8027..8046)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8187..8209
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8203..8394)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(9706..10788)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     10750..10769
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(10789..10866)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
       61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
      121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
      181 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
      241 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat ataccatgga
      301 ctacaaagac catgacggtg attataaaga tcatgacatc gattacaagg atgacgatga
      361 caagatggcc cccaagaaga agaggaaggt gggccgcgga atggaaaaaa aggaatttcg
      421 tgttttgata aaatactgtt ttctgaaggg aaaaaataca gtggaagcaa aaacttggct
      481 tgataatgag tttccggact ctgccccagg gaaatcaaca ataattgatt ggtatgcaaa
      541 attcaagcgt ggtgaaatga gcacggagga cggtgaacgc agtggacgcc cgaaagaggt
      601 ggttaccgac gaaaacatca aaaaaatcca caaaatgatt ttgaatgacc gtaaaatgaa
      661 gttgatcgag atagcagagg ccttaaagat atcaaaggaa cgtgttggtc atatcattca
      721 tcaatatttg gatatgcgga agctctgtgc gaaatgggtg ccgcgcgagc tcacatttga
      781 ccaaaaacaa cgacgtgttg atgattctaa gcggtgtttg cagctgttaa ctcgtaatac
      841 acccgagttt ttccgtcgat atgtgacaat ggatgaaaca tggctccatc actacactcc
      901 tgagtccaat cgacagtcgg ctgagtggac agcgaccggt gaaccgtctc cgaagcgtgg
      961 aaagactcaa aagtccgctg gcaaagtaat ggcctctgtt ttttgggatg cgcatggaat
     1021 aatttttatc gattatcttg agaagggaaa aaccatcaac agtgactatt atatggcgtt
     1081 attggagcgt ttgaaggtcg aaatcgcggc aaaacggccc cacatgaaga agaaaaaagt
     1141 gttgttccac caagacaacg caccgtgcca caagtcattg agaacgatgg caaaaattca
     1201 tgaattgggc ttcgaattgc ttccccaccc gccgtattct ccagatctgg cccccagcga
     1261 ctttttcttg ttctcagacc tcaaaaggat gctcgcaggg aaaaaatttg gctgcaatga
     1321 agaggtgatc gccgaaactg aggcctattt tgaggcaaaa ccgaaggagt actaccaaaa
     1381 tggtatcaaa aaattggaag gtcgttataa tcgttgtatc gctcttgaag ggaactatgt
     1441 tgaaagcggt tccgaaactc ccggtacatc agaaagcgcg acccccgaaa gcatggataa
     1501 aaagtattct attggtttag ctatcggcac aaatagcgtc ggatgggcgg tgatcactga
     1561 tgaatataag gttccgtcta aaaagttcaa ggttctggga aatacagacc gccacagtat
     1621 caaaaaaaat cttatagggg ctcttttatt tgacagtgga gagacagcgg aagcgactcg
     1681 tctcaaacgg acagctcgta gaaggtatac acgtcggaag aatcgtattt gttatctaca
     1741 ggagattttt tcaaatgaga tggcgaaagt agatgatagt ttctttcatc gacttgaaga
     1801 gtcttttttg gtggaagaag acaagaagca tgaacgtcat cctatttttg gaaatatagt
     1861 agatgaagtt gcttatcatg agaaatatcc aactatctat catctgcgaa aaaaattggt
     1921 agattctact gataaagcgg atttgcgctt aatctatttg gccttagcgc atatgattaa
     1981 gtttcgtggt cattttttga ttgagggaga tttaaatcct gataatagtg atgtggacaa
     2041 actatttatc cagttggtac aaacctacaa tcaattattt gaagaaaacc ctattaacgc
     2101 aagtggagta gatgctaaag cgattctttc tgcacgattg agtaaatcaa gacgattaga
     2161 aaatctcatt gctcagctcc ccggtgagaa gaaaaatggc ttatttggga atctcattgc
     2221 tttgtcattg ggtttgaccc ctaattttaa atcaaatttt gatttggcag aagatgctaa
     2281 attacagctt tcaaaagata cttacgatga tgatttagat aatttattgg cgcaaattgg
     2341 agatcaatat gctgatttgt ttttggcagc taagaattta tcagatgcta ttttactttc
     2401 agatatccta agagtaaata ctgaaataac taaggctccc ctatcagcat caatgattaa
     2461 acgctacgat gaacatcatc aagacttgac tcttttaaaa gctttagttc gacaacaact
     2521 tccagaaaag tataaagaaa tcttttttga tcaatcaaaa aacggatatg caggttatat
     2581 tgatggggga gctagccaag aagaatttta taaatttatc aaaccaattt tagaaaaaat
     2641 ggatggtact gaggaattat tggtgaaact aaatcgtgaa gatttgctgc gcaagcaacg
     2701 gacctttgac aacggctcta ttccccatca aattcacttg ggtgagctgc atgctatttt
     2761 gagaagacaa gaagactttt atccattttt aaaagacaat cgtgagaaga ttgaaaaaat
     2821 cttgactttt cgaattcctt attatgttgg tccattggcg cgtggcaata gtcgttttgc
     2881 atggatgact cggaagtctg aagaaacaat taccccatgg aattttgaag aagttgtcga
     2941 taaaggtgct tcagctcaat catttattga acgcatgaca aactttgata aaaatcttcc
     3001 aaatgaaaaa gtactaccaa aacatagttt gctttatgag tattttacgg tttataacga
     3061 attgacaaag gtcaaatatg ttactgaagg aatgcgaaaa ccagcatttc tttcaggtga
     3121 acagaagaaa gccattgttg atttactctt caaaacaaat cgaaaagtaa ccgttaagca
     3181 attaaaagaa gattatttca aaaaaataga atgttttgat agtgttgaaa tttcaggagt
     3241 tgaagataga tttaatgctt cattaggtac ctaccatgat ttgctaaaaa ttattaaaga
     3301 taaagatttt ttggataatg aagaaaatga agatatctta gaggatattg ttttaacatt
     3361 gaccttattt gaagataggg agatgattga ggaaagactt aaaacatatg ctcacctctt
     3421 tgatgataag gtgatgaaac agcttaaacg tcgccgttat actggttggg gacgtttgtc
     3481 tcgaaaattg attaatggta ttagggataa gcaatctggc aaaacaatat tagatttttt
     3541 gaaatcagat ggttttgcca atcgcaattt tatgcagctg atccatgatg atagtttgac
     3601 atttaaagaa gacattcaaa aagcacaagt gtctggacaa ggcgatagtt tacatgaaca
     3661 tattgcaaat ttagctggta gccctgctat taaaaaaggt attttacaga ctgtaaaagt
     3721 tgttgatgaa ttggtcaaag taatggggcg gcataagcca gaaaatatcg ttattgaaat
     3781 ggcacgtgaa aatcagacaa ctcaaaaggg ccagaaaaat tcgcgagagc gtatgaaacg
     3841 aatcgaagaa ggtatcaaag aattaggaag tcagattctt aaagagcatc ctgttgaaaa
     3901 tactcaattg caaaatgaaa agctctatct ctattatctc caaaatggaa gagacatgta
     3961 tgtggaccaa gaattagata ttaatcgttt aagtgattat gatgtcgatg ccattgttcc
     4021 acaaagtttc cttaaagacg attcaataga caataaggtc ttaacgcgtt ctgataaaaa
     4081 tcgtggtaaa tcggataacg ttccaagtga agaagtagtc aaaaagatga aaaactattg
     4141 gagacaactt ctaaacgcca agttaatcac tcaacgtaag tttgataatt taacgaaagc
     4201 tgaacgtgga ggtttgagtg aacttgataa agctggtttt atcaaacgcc aattggttga
     4261 aactcgccaa atcactaagc atgtggcaca aattttggat agtcgcatga atactaaata
     4321 cgatgaaaat gataaactta ttcgagaggt taaagtgatt accttaaaat ctaaattagt
     4381 ttctgacttc cgaaaagatt tccaattcta taaagtacgt gagattaaca attaccatca
     4441 tgcccatgat gcgtatctaa atgccgtcgt tggaactgct ttgattaaga aatatccaaa
     4501 acttgaatcg gagtttgtct atggtgatta taaagtttat gatgttcgta aaatgattgc
     4561 taagtctgag caagaaatag gcaaagcaac cgcaaaatat ttcttttact ctaatatcat
     4621 gaacttcttc aaaacagaaa ttacacttgc aaatggagag attcgcaaac gccctctaat
     4681 cgaaactaat ggggaaactg gagaaattgt ctgggataaa gggcgagatt ttgccacagt
     4741 gcgcaaagta ttgtccatgc cccaagtcaa tattgtcaag aaaacagaag tacagacagg
     4801 cggattctcc aaggagtcaa ttttaccaaa aagaaattcg gacaagctta ttgctcgtaa
     4861 aaaagactgg gatccaaaaa aatatggtgg ttttgatagt ccaacggtag cttattcagt
     4921 cctagtggtt gctaaggtgg aaaaagggaa atcgaagaag ttaaaatccg ttaaagagtt
     4981 actagggatc acaattatgg aaagaagttc ctttgaaaaa aatccgattg actttttaga
     5041 agctaaagga tataaggaag ttaaaaaaga cttaatcatt aaactaccta aatatagtct
     5101 ttttgagtta gaaaacggtc gtaaacggat gctggctagt gccggagaat tacaaaaagg
     5161 aaatgagctg gctctgccaa gcaaatatgt gaatttttta tatttagcta gtcattatga
     5221 aaagttgaag ggtagtccag aagataacga acaaaaacaa ttgtttgtgg agcagcataa
     5281 gcattattta gatgagatta ttgagcaaat cagtgaattt tctaagcgtg ttattttagc
     5341 agatgccaat ttagataaag ttcttagtgc atataacaaa catagagaca aaccaatacg
     5401 tgaacaagca gaaaatatta ttcatttatt tacgttgacg aatcttggag ctcccgctgc
     5461 ttttaaatat tttgatacaa caattgatcg taaacgatat acgtctacaa aagaagtttt
     5521 agatgccact cttatccatc aatccatcac tggtctttat gaaacacgca ttgatttgag
     5581 tcagctagga ggtgaccacc accaccacca ccactgagat ccggctgcta acaaagcccg
     5641 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     5701 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gatatcccgc
     5761 aagaggcccg gcagtaccgg cataaccaag cctatgccta cagcatccag ggtgacggtg
     5821 ccgaggatga cgatgagcgc attgttagat ttcatacacg gtgcctgact gcgttagcaa
     5881 tttaactgtg ataaactacc gcattaaagc tagcttatcg atgataagct gtcaaacatg
     5941 agaattaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     6001 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     6061 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     6121 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     6181 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6241 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6301 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6361 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     6421 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6481 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6541 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6601 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6661 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     6721 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6781 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6841 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6901 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6961 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     7021 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     7081 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     7141 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     7201 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7261 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7321 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7381 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7441 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7501 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7561 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7621 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7681 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7741 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7801 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7861 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     7921 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     7981 cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg
     8041 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     8101 gccccgacac ccaccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     8161 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     8221 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     8281 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     8341 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     8401 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     8461 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     8521 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg
     8581 cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat
     8641 ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa
     8701 accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca
     8761 cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag
     8821 ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggccaggacc caacgctgcc
     8881 cgagatgcgc cgcgtgcggc tgctggagat ggcggacgcg atggatatgt tctgccaagg
     8941 gttggtttgc gcattcacag ttctccgcaa gaattgattg gctccaattc ttggagtggt
     9001 gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg gctccatgca
     9061 ccgcgacgca acgcggggag gcagacaagg tatagggcgg cgcctacaat ccatgccaac
     9121 ccgttccatg tgctcgccga ggcggcataa atcgccgtga cgatcagcgg tccaatgatc
     9181 gaagttaggc tggtaagagc cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct
     9241 acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga
     9301 atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac gtagcccagc
     9361 gcgtcggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
     9421 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     9481 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     9541 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     9601 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     9661 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     9721 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     9781 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     9841 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     9901 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     9961 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
    10021 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
    10081 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
    10141 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
    10201 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
    10261 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
    10321 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
    10381 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
    10441 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
    10501 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
    10561 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
    10621 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
    10681 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
    10741 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
    10801 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
    10861 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
    10921 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatg
//
